CN102329827A - Sheep placenta amino acid and low molecular weight polypeptide, and preparation method for sheep placenta amino acid and low molecular weight polypeptide - Google Patents
Sheep placenta amino acid and low molecular weight polypeptide, and preparation method for sheep placenta amino acid and low molecular weight polypeptide Download PDFInfo
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- CN102329827A CN102329827A CN201110193935A CN201110193935A CN102329827A CN 102329827 A CN102329827 A CN 102329827A CN 201110193935 A CN201110193935 A CN 201110193935A CN 201110193935 A CN201110193935 A CN 201110193935A CN 102329827 A CN102329827 A CN 102329827A
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Abstract
The invention relates to a method for preparing sheep embryo and sheep placenta biological products, in particular to a method for preparing sheep placenta amino acid and low molecular weight polypeptide. The method comprises the following steps of: 1) pretreating sheep embryo and sheep placenta raw materials; 2) performing ultra-fine pulverization and cell wall breaking of sheep embryo and sheep placenta; 3) performing extraction and enzymolysis on the sheep embryo and the sheep placenta; and 4) performing fractional refining on a stock solution of the sheep embryo and the sheep placenta obtained in the step 3) by using a sephadex chromatography method according to molecular weight to obtain the sheep placenta amino acid and the low molecular weight polypeptide. By the sheep placenta amino acid and the low molecular weight polypeptide prepared by the method, active materials of the sheep embryo and the sheep placenta are retained furthest and are easily absorbed by a human body comprehensively; and the process is advanced and reasonable, and the quality is controllable.
Description
Technical field
The present invention relates to a kind of sheep embryo and Placenta caprae seu ovis biological products and preparation method thereof.
Background technology
China utilizes that various organs, tissue and the meta-bolites of animal are cured the disease, diseases prevention and health care with a long history, existing complete pharmacy ancient books and records---" legendary god of farming's book on Chinese herbal medicine opinion " just has the record of multiple animal pharmaceuticals such as cow-bezoar, rhinoceros horn, pilose antler, donkey-hide gelatin.Explained that China utilizes animal pharmaceuticals to process, extract already, refiningly forms comparatively purified medicament on pharmaceutical technology.
Placenta (Placenta) has another name called " Placenta Hominis "; Be conventional Chinese medicine; Modern medicine study shows: placenta contains biologically active substance such as panimmunity sphaeroprotein, bioactive peptide and amino acid, mineral matter and other components (the general designation viable cell is plain), is a kind of regulator and nutrition, cosmetic health product of body's immunity.Scientist compares the experiment back to multiple animal placenta and finds that the cell tissue structure of Placenta caprae seu ovis is the most similar with people's placenta, is expected to become with the human placenta to have the surrogate that equal health is worth.
In recent years, occurred two kinds of novel Placenta caprae seu ovis biological products of animal placenta hydrolyzed solution and placenta vat liquor at home and abroad again in succession,, caused gazing at of people because of its unique usefulness.But we think that above-mentioned two kinds of preparations respectively have drawback, and the former destroys the natural active matter in the placenta in hydrolytic process, loses the meaning of utilizing placenta; Though the latter has multiple natural active matter, polypeptide and aminoacids content are very little, and the two is combined together, have both kept the natural active matter in the placenta, increase its polypeptide and aminoacids content again.This is a new breakthrough in sheep placenta and the development research of Placenta caprae seu ovis biological products undoubtedly.
Summary of the invention
In order to solve above-mentioned technical problem, an object of the present invention is to provide the preparation method of a kind of sheep placental extract amino acid and low molecular polypeptide.Another object of the present invention provides above-mentioned sheep placental extract amino acid and low molecular polypeptide.
In order to realize first above-mentioned purpose, the technical scheme below the present invention has adopted:
The preparation method of a kind of sheep placental extract amino acid and low molecular polypeptide, this method comprises the steps:
1) sheep embryo and the pre-treatment of Placenta caprae seu ovis raw material: getting sheep embryo and Placenta caprae seu ovis is raw material, cleans, and removes impurity, is ground into meal under 0～5 ℃ of low temperature;
2) sheep embryo and Placenta caprae seu ovis micronizing, cell wall breaking: behind the freezing 1～3h of sheep embryo and Placenta caprae seu ovis meal low temperature-40 ℃～-50 ℃ ,-30～-10 ℃ of micronizing, cell wall breaking reaches 0.1～10 μ m granularity;
3) extraction of sheep embryo and Placenta caprae seu ovis and enzymolysis: sheep embryo behind the cell wall breaking and Placenta caprae seu ovis slurries add 3～8 times of distillation water temperature lixiviates and get after-filtration must filtrate A and precipitate A; Suction filtration got hydrolyzed solution B after precipitate A adopted protease hydrolyzed; Merging filtrate A and hydrolyzed solution B, it is plain to get the plain refining sheep placenta of sheep embryo and Placenta caprae seu ovis;
4) adopt the sephadex chromatography method; According to the molecular weight size to above-mentioned steps 3) in the sheep placenta that obtains and the plain stoste of Placenta caprae seu ovis to carry out classification refining; Stationary phase is Sephadex G-10, and water, 2%NaCl eluant solution are collected amino acid and the molecular weight < different components of 700 polypeptide successively.
As preferably, sheep embryo and Placenta caprae seu ovis with 2～3 monthly ages in the above-mentioned step 1) are raw material.
As preferably, proteolytic enzyme adopts that one or both mix in papoid and the trypsinase in the above-mentioned step 3).
As preferably, the usage quantity of proteolytic enzyme is 0.1%～2% of a precipitate A weight in the above-mentioned step 3).
Preferred as again, the usage quantity of proteolytic enzyme is 0.3%～1% of a precipitate A weight in the above-mentioned step 3).
As preferably, proteolytic enzyme adopts papoid to mix for two kinds with trypsinase in the above-mentioned step 3), and the temperature of enzyme digestion reaction is 40～50 ℃, and pH is 7.0～8.0, papoid and tryptic enzyme proportioning 1:0.5～2.
Preferred as again, the temperature of enzyme digestion reaction is 45 ℃ in the step 3), and pH is 7.5, papoid and tryptic enzyme proportioning 1:1.
In order to realize second above-mentioned purpose, sheep placental extract amino acid and low molecular polypeptide that the present invention also provides above-mentioned any technical scheme to prepare.
The present invention is owing to adopted above-mentioned technical scheme; This technological process of manufacture can fully extract the effective constituent in sheep placenta and the Placenta caprae seu ovis, and the active substance of sheep placenta and Placenta caprae seu ovis is maximized the retention, and is convenient to human body again and absorbs comprehensively; Technology is advanced, reasonable, quality controllable.Adopt aforesaid method to make sheep placental extract amino acid and low molecular polypeptide, have following effect:
1, regulate female incretion, improve the human body skin situation fully and effectively, beautifying face and moistering lotion, skin whitening strengthens skin elasticity, removes blackspot, prevents wrinkle, striae of pregnancy and delays senility;
2, can prevent infertility, miscarriage, premature labor;
3, the smart marrow of mending of benefit increases the production of man's sperm, improves Infertility (men and women is all effective), increases sexual function;
4, adjustment hormone increases the blood manufacturing, improves anaemia, the deficiency of vital energy, strengthens cell viability, quickens postpartum and recovers, and helps cesarean lactation;
5, improve the human immunologic function, postpone Women, improve climacteric syndrome;
6, promote breast development, make bagged breast recovery in postpartum strong, high resilience;
7, postoperative physical recovery;
8, antianaphylaxis, antitumor.
Embodiment
Embodiment 1
The preparation method of a kind of sheep placental extract amino acid and low molecular polypeptide, this method comprises the steps:
1) sheep embryo and the pre-treatment of Placenta caprae seu ovis raw material: sheep embryo and Placenta caprae seu ovis with 2-3 monthly age are raw material, clean, remove impurity, are ground into meal under the 0-5 ℃ of low temperature.
2) sheep embryo and Placenta caprae seu ovis micronizing, cell wall breaking: with low temperature (20 ℃) micronizing (the medicinal supper micron mill of WZJ100) behind sheep embryo and Placenta caprae seu ovis meal freezing (40 ℃～-45 ℃) 2h, cell wall breaking reaches 0.1-10 μ m granularity.
3) extraction of sheep embryo and Placenta caprae seu ovis and enzymolysis: sheep embryo behind micronizing, cell wall breaking and Placenta caprae seu ovis slurries add 6 times of distillation water temperature lixiviates and get after-filtration must filtrate A and precipitate A; Precipitate A with papoid and trypsin digestion after suction filtration get hydrolyzed solution B; 45 ℃ of hydrolysis temperatures; Enzyme proportioning 1g:1g, pH7.5, enzyme-to-substrate are 20mg:3g; Merging filtrate A and hydrolyzed solution B, it is plain to get the plain refining sheep placenta of sheep embryo and Placenta caprae seu ovis.
Adopt polydextran gel (DEAE SepharoseTM Fast Flow, Sephadex G-50) according to the molecular weight size to above-mentioned steps 3) in the sheep placenta that obtains and the plain stoste of Placenta caprae seu ovis to carry out classification refining, the refining sheep placenta that obtains amino acid, micromolecule polypeptide and various molecular weight proteins is plain.Its technical process is following:
1. amino acid separation, molecular weight 700 polypeptide: appearance on the plain refining sheep placenta element of sheep embryo and Placenta caprae seu ovis, stationary phase Sephadex G-10, water successively, the 2%NaCl eluant solution is collected different components.
2. the polypeptide of isolated molecule amount 1000-5000: appearance on the water elution liquid 1., stationary phase Sephadex G-25, water successively, 0%-2%NaCl solution gradient wash-out is collected different components.
3. the polypeptide of isolated molecule amount 5000-30000: appearance on the water elution liquid 2., stationary phase Sephadex G-50, water successively, 0%-2%NaCl solution gradient wash-out is collected different components.
4. the protein of isolated molecule amount 30000-150000: appearance on the water elution liquid 3., stationary phase Sephadex G-100, water successively, 0%-2%NaCl solution gradient wash-out is collected different components.
5. the protein of isolated molecule amount 150000-600000: appearance on the water elution liquid 4., stationary phase Sephadex G-200, water successively, 0%-2%NaCl solution gradient wash-out is collected different components.
Adopt gel permeation chromatography-evaporat light scattering (GPC-ELSD) method to measure the MWD of sheep placenta and Placenta caprae seu ovis biological products, formulate its quality standard.
Step is following:
1. it is plain to get sheep embryo and Placenta caprae seu ovis stoste, refined powder or refining sheep placenta, adds an amount of pure water heating or ultrasonic dissolution, puts coldly, and constant volume filters, and gets subsequent filtrate as need testing solution;
2. liquid-phase condition: moving phase is pure water, flow velocity 1ml/min, 30 ℃ of column temperatures;
3. stationary phase: gel filtration chromatography post (GPC) chromatographic column Shodex OHpak SB-806 HQ, Shodex OHpak SB-805 HQ, Shodex OHpak SB-804 HQ, Shodex OHpak SB-803 HQ series connection;
4. detector: light scattering detector (ELSD), vaporization temperature: 100 ℃, atomization temperature: 80 ℃, nitrogen flow rate: 1.5L/min.
Embodiment 2
The preparation method of a kind of sheep placental extract amino acid and low molecular polypeptide, this method enzymolysis in step 3) adopts the papoid, and other technologies scheme such as embodiment 1 are said.
Claims (9)
1. the preparation method of sheep placental extract amino acid and low molecular polypeptide is characterized in that this method comprises the steps:
1) sheep embryo and the pre-treatment of Placenta caprae seu ovis raw material: getting sheep embryo and Placenta caprae seu ovis is raw material, cleans, and removes impurity, is ground into meal under 0～5 ℃ of low temperature;
2) sheep embryo and Placenta caprae seu ovis micronizing, cell wall breaking: behind the freezing 1～3h of sheep embryo and Placenta caprae seu ovis meal low temperature-40 ℃～-50 ℃ ,-30～-10 ℃ of micronizing, cell wall breaking reaches 0.1～10 μ m granularity;
3) extraction of sheep embryo and Placenta caprae seu ovis and enzymolysis: sheep embryo behind the cell wall breaking and Placenta caprae seu ovis slurries add 3～10 times of distillation water temperature lixiviates and get after-filtration must filtrate A and precipitate A; Suction filtration got hydrolyzed solution B after precipitate A adopted protease hydrolyzed; Merging filtrate A and hydrolyzed solution B, it is plain to get the plain refining sheep placenta of sheep embryo and Placenta caprae seu ovis;
4) adopt the sephadex chromatography method; According to the molecular weight size to above-mentioned steps 3) in the sheep placenta that obtains and the plain stoste of Placenta caprae seu ovis to carry out classification refining; Stationary phase is Sephadex G-10; Water, 2%NaCl eluant solution are collected amino acid and polypeptide and the molecular weight < different components of 700 polypeptide successively.
2. the preparation method of sheep placental extract amino acid according to claim 1 and low molecular polypeptide is characterized in that: sheep embryo and Placenta caprae seu ovis with 2～3 monthly ages in the step 1) are raw material.
3. the preparation method of sheep placental extract amino acid according to claim 1 and low molecular polypeptide is characterized in that: proteolytic enzyme adopts that one or both mix in papoid and the trypsinase in the step 3).
4. the preparation method of sheep placental extract amino acid according to claim 1 and low molecular polypeptide is characterized in that: the usage quantity of proteolytic enzyme is 0.1%～2% of a precipitate A weight in the step 3).
5. the preparation method of sheep placental extract amino acid according to claim 4 and low molecular polypeptide is characterized in that: the usage quantity of proteolytic enzyme is 0.3%～1% of a precipitate A weight in the step 3).
6. according to the preparation method of claim 1 or 2 or 3 or 4 or 5 described sheep placental extract amino acid and low molecular polypeptide; It is characterized in that: proteolytic enzyme adopts papoid to mix for two kinds with trypsinase in the step 3); The temperature of enzyme digestion reaction is 40～50 ℃; PH is 7.0～8.0, papoid and tryptic enzyme proportioning 1:0.5～2.
7. the preparation method of sheep placental extract amino acid according to claim 6 and low molecular polypeptide is characterized in that: the temperature of enzyme digestion reaction is 45 ℃ in the step 3), and pH is 7.5, papoid and tryptic enzyme proportioning 1:1.
8. the sheep placental extract amino acid and the low molecular polypeptide that prepare according to claim 1 or 2 or 3 or 4 or 5 described methods.
9. sheep placental extract amino acid and low molecular polypeptide that method according to claim 6 prepares.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104398399A (en) * | 2014-11-14 | 2015-03-11 | 唯美度科技（北京）有限公司 | Composite glycopeptide breast beauty cream and preparation method thereof |
| CN104606668A (en) * | 2015-02-06 | 2015-05-13 | 深圳唯美度生物科技有限公司 | Breast enlargement gel containing complex polypeptides and process thereof |
| CN107184609A (en) * | 2017-06-09 | 2017-09-22 | 郑州源创吉因实业有限公司 | A kind of lossless sterile bulk preparation method of Human plactnta |
| CN108396051A (en) * | 2018-03-28 | 2018-08-14 | 通化百泉保健食品有限公司 | A kind of industrialized producing technology of sheep placenta lamb albumen oligopeptide |
| CN108866136A (en) * | 2018-08-01 | 2018-11-23 | 李文锦 | A kind of preparation method of low molecular polypeptide |
| CN109971810A (en) * | 2019-04-10 | 2019-07-05 | 长沙嘉兰生物科技有限公司 | A kind of sheep embryo extract protein small peptide and its preparation process |
| CN111647039A (en) * | 2020-07-07 | 2020-09-11 | 肖启森 | Method for extracting sheep embryo element active peptide |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104398399A (en) * | 2014-11-14 | 2015-03-11 | 唯美度科技（北京）有限公司 | Composite glycopeptide breast beauty cream and preparation method thereof |
| CN104606668A (en) * | 2015-02-06 | 2015-05-13 | 深圳唯美度生物科技有限公司 | Breast enlargement gel containing complex polypeptides and process thereof |
| CN107184609A (en) * | 2017-06-09 | 2017-09-22 | 郑州源创吉因实业有限公司 | A kind of lossless sterile bulk preparation method of Human plactnta |
| CN108396051A (en) * | 2018-03-28 | 2018-08-14 | 通化百泉保健食品有限公司 | A kind of industrialized producing technology of sheep placenta lamb albumen oligopeptide |
| CN108866136A (en) * | 2018-08-01 | 2018-11-23 | 李文锦 | A kind of preparation method of low molecular polypeptide |
| CN109971810A (en) * | 2019-04-10 | 2019-07-05 | 长沙嘉兰生物科技有限公司 | A kind of sheep embryo extract protein small peptide and its preparation process |
| CN111647039A (en) * | 2020-07-07 | 2020-09-11 | 肖启森 | Method for extracting sheep embryo element active peptide |
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